Urinary genome detection and tracking of Hantaan virus from hemorrhagic fever with renal syndrome patients using multiplex PCR-based next-generation sequencing

PLoS Negl Trop Dis. 2021 Sep 28;15(9):e0009707. doi: 10.1371/journal.pntd.0009707. eCollection 2021 Sep.

Abstract

Background: Hantavirus infection occurs through the inhalation of aerosolized excreta, including urine, feces, and saliva of infected rodents. The presence of Hantaan virus (HTNV) RNA or infectious particles in urine specimens of patient with hemorrhagic fever with renal syndrome (HFRS) remains to be investigated.

Methodology/principal findings: We collected four urine and serum specimens of Republic of Korea Army (ROKA) patients with HFRS. We performed multiplex PCR-based next-generation sequencing (NGS) to obtain the genome sequences of clinical HTNV in urine specimens containing ultra-low amounts of viral genomes. The epidemiological and phylogenetic analyses of HTNV demonstrated geographically homogenous clustering with those in Apodemus agrarius captured in highly endemic areas, indicating that phylogeographic tracing of HTNV genomes reveals the potential infection sites of patients with HFRS. Genetic exchange analyses showed a genetic configuration compatible with HTNV L segment exchange in nature.

Conclusion/significance: Our results suggest that whole or partial genome sequences of HTNV from the urine enabled to track the putative infection sites of patients with HFRS by phylogeographically linking to the zoonotic HTNV from the reservoir host captured at endemic regions. This report raises awareness among physicians for the presence of HTNV in the urine of patients with HFRS.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Genome, Viral*
  • Hantaan virus / classification
  • Hantaan virus / genetics
  • Hantaan virus / isolation & purification*
  • Hemorrhagic Fever with Renal Syndrome / urine
  • Hemorrhagic Fever with Renal Syndrome / virology*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Multiplex Polymerase Chain Reaction
  • Phylogeny
  • Republic of Korea
  • Urine / virology*

Grants and funding

This work was supported by the Agency for Defense Development (UE202026GD to J.-W.S.), the Research Program To Solve Social Issues of the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (NRF-2017M3A9E4061992 to J.-W.S.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.